In vitro regulation of pericellular proteolysis in prostatic tumor cells treated with bombesin.
Bombesin is a potent inducer of signal trasduction pathways involved in the proliferation and invasion of androgen-insensitive prostatic tumor cells. This study examines the bombesin-mediated modulation of pericellular proteolysis, monitoring cell capability to migrate and invade basement membranes, using a chemo-invasion assay and analyzing protease production. The results suggest that bombesin could modulate the invasive potential of prostatic cell lines regulating secretion and cell-surface uptake of uPA and MMP-9 activation. In fact, in PC3 and DU145 cells but not in LNCaP cells, urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) are induced by bombesin treatment. Bombesin also stimulates cell proliferation and this effect can be inhibited blocking uPA by antibodies and/or uPA inhibitor p-aminobenzamidine. Moreover, HMW-uPA induces cell proliferation in LNCaP cells, which do not produce uPA in the basal conditions, while PC3 and DU145 cell growth is supported by autocrine production of uPA. The increment of uPA activity on the external plasma membrane causes an increased pericellular plasmin activation. This effect is inhibited by antibodies against uPA and by p-aminobenzamidine. Similarly to EGF, bombesin stimulates secretion and activation of MMP-9 and TIMP-1 production. MMP-9 activation can be also obtained by HMW-uPA treatment, suggesting that plasma-membrane-bound uPA can start a proteolytic cascade involving MMP-9. Therefore, in in vitro assays, bombesin is able to modulate pericellular proteolysis and cell proliferation, differently distributing and activating proteolytic activities. This effect can be related to the "non-random" degradation of the extracellular matrix in which membrane uPA-uPAreceptor complexes could start bombesin-induced directional protein degradation during metastatic spread.
Festuccia
C
C
Dipartimento di Medicina Sperimentale, Università dell'Aquila, L'Aquila, Italy.
Guerra
F
F
D'Ascenzo
S
S
Giunciuglio
D
D
Albini
A
A
Bologna
M
M
D000906:Antibodies / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001550:Benzamidines / Q000494:pharmacology; D001839:Bombesin / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002462:Cell Membrane / Q000201:enzymology; D003094:Collagen; D017364:Collagenases / Q000096:biosynthesis; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D004789:Enzyme Activation; D004790:Enzyme Induction; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005341:Fibrinolysin / Q000378:metabolism; D019886:Gastrin-Releasing Peptide / Q000494:pharmacology; D006801:Humans; D007797:Laminin; D008297:Male; D020780:Matrix Metalloproteinase 9; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000187:drug effects* / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000473:pathology; D011509:Proteoglycans; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism
D000906:Antibodies; D000970:Antineoplastic Agents; D001550:Benzamidines; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D007797:Laminin; D017395:Plasminogen Activator Inhibitor 1; D011509:Proteoglycans; D015842:Serine Proteinase Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1; C059485:matrigel; D004815:Epidermal Growth Factor; C006500:4-aminobenzamidine; D019886:Gastrin-Releasing Peptide; D003094:Collagen; D010447:Peptide Hydrolases; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; D017364:Collagenases; D020780:Matrix Metalloproteinase 9; D001839:Bombesin